LOCUS       sequence-450927-        7091 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7091)
  AUTHORS   Calvisi DF, Wang C, Ho C, Ladu S, Lee SA, Mattu S, Destefanis G, 
            Delogu S, Zimmermann A, Ericsson J, Brozzetti S, Staniscia T, Chen 
            X, Dombrowski F, Evert M
  TITLE     Increased lipogenesis, induced by AKT-mTORC1-RPS6 signaling, 
            promotes development of human hepatocellular carcinoma.
  JOURNAL   Gastroenterology. 2011 Mar;140(3):1071-83. Epub 2010 Dec 11.
  PUBMED    21147110
REFERENCE   2  (bases 1 to 7091)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Gastroenterology."; date: "2011-03"; volume: "140(3)"; pages: 
            "1071-83. Epub 2010 Dec 11"
COMMENT     Sequence Label: pT3-myr-AKT-HA
FEATURES             Location/Qualifiers
     source          1..7091
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1417..1436)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    1489..1522
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1663..2841
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1894..2832
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2789..2809
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     2858..2877
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2858..2876
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2955..2979
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3000..3020
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSK"
     CDS             4473..4499
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(4519..4543)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4596..4637
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(4675..4692)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4681..4905
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    4975..5008
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     5064..5084
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(5466..5483)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5466..5482)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5475..5497)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5691..5710)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5810..5832
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5870..5888)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5956..6060
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6061..6129,6130..6921)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6279..6298)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg catgcctgca
     1021 ggtcgactct agacctatac agttgaagtc ggaagtttac atacacttaa gttggagtca
     1081 ttaaaactcg tttttcaact actccacaaa tttcttgtta acaaacaata gttttggcaa
     1141 gtcagttagg acatctactt tgtgcatgac acaagtcatt tttccaacaa ttgtttacag
     1201 acagattatt tcacttataa ttcactgtat cacaattcca gtgggtcaga agtttacata
     1261 cactaagttg actgtgcctt taaacagctt ggaaaattcc agaaaatgat gtcatggctt
     1321 tagaagcttc tgatagacta attgacatca tttgagtcaa ttggaggtgt acctgtggat
     1381 gtatttcaag gggatccaga catgataaga tacattgatg agtttggaca aaccacaact
     1441 agaatgcagc ggccgctagc tcgagatctt aattaaacta gtatcgatat aacttcgtat
     1501 aatgtatgct atacgaagtt atatgtacgg gccagatata cgcgttgaca ttgattattg
     1561 actaggcttt tgcaaaaagc tttgcaaaga tggataaagt tttaaacaga gaggaatctt
     1621 tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa ttggctccgg tgcccgtcag
     1681 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
     1741 accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
     1801 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
     1861 ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg
     1921 cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac ctggctgcag
     1981 tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt gggagagttc gaggccttgc
     2041 gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct ggcctgggcg ctggggccgc
     2101 cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa gtctctagcc
     2161 atttaaaatt tttgatgacc tgctgcgacg ctttttttct ggcaagatag tcttgtaaat
     2221 gcgggccaag atctgcacac tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc
     2281 cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc accgagaatc
     2341 ggacgggggt agtctcaagc tggccggcct gctctggtgc ctggcctcgc gccgccgtgt
     2401 atcgccccgc cctgggcggc aaggctggcc cggtcggcac cagttgcgtg agcggaaaga
     2461 tggccgcttc ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg ctcgggagag
     2521 cgggcgggtg agtcacccac acaaaggaaa agggcctttc cgtcctcagc cgtcgcttca
     2581 tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg attagttctc gagcttttgg
     2641 agtacgtcgt ctttaggttg gggggagggg ttttatgcga tggagtttcc ccacactgag
     2701 tgggtggaga ctgaagttag gccagcttgg cacttgatgt aattctcctt ggaatttgcc
     2761 ctttttgagt ttggatcttg gttcattctc aagcctcaga cagtggttca aagttttttt
     2821 cttccatttc aggtgtcgtg aggaattagc ttggtactaa tacgactcac tatagggaga
     2881 cccaagctgg ctaggtaagc ttggtaccga gctcggatcc actagtccag tgtggtggaa
     2941 ttctgcagat atcaacaagt ttgtacaaaa aagcaggctc cgcggccgcc cccttcacca
     3001 tggggagcag caagagcaag cccaagtcta gaatgaacga cgtagccatt gtgaaggagg
     3061 gctggctgca caaacgaggg gaatatatta aaacctggcg gccacgctac ttcctcctca
     3121 agaacgatgg cacctttatt ggctacaagg aacggcctca ggatgtggat cagcgagagt
     3181 ccccactcaa caacttctca gtggcacaat gccagctgat gaagacagag cggccaaggc
     3241 ccaacacctt tatcatccgc tgcctgcagt ggaccacagt cattgagcgc accttccatg
     3301 tggaaacgcc tgaggagcgg gaagaatggg ccaccgccat tcagactgtg gccgatggac
     3361 tcaagaggca ggaagaagag acgatggact tccgatcagg ctcacccagt gacaactcag
     3421 gggctgaaga gatggaggtg tccctggcca agcccaagca ccgtgtgacc atgaacgagt
     3481 ttgagtacct gaaactactg ggcaagggca cctttgggaa agtgattctg gtgaaagaga
     3541 aggccacagg ccgctactat gccatgaaga tcctcaagaa ggaggtcatc gtcgccaagg
     3601 atgaggttgc ccacacgctt actgagaacc gtgtcctgca gaactctagg catcccttcc
     3661 ttacggccct caagtactca ttccagaccc acgaccgcct ctgctttgtc atggagtatg
     3721 ccaacggggg cgagctcttc ttccacctgt ctcgagagcg cgtgttctcc gaggaccggg
     3781 cccgcttcta tggtgcggag attgtgtctg ccctggacta cttgcactcc gagaagaacg
     3841 tggtgtaccg ggacctgaag ctggagaacc tcatgctgga caaggacggg cacatcaaga
     3901 taacggactt cgggctgtgc aaggagggga tcaaggatgg tgccactatg aagacattct
     3961 gcggaacgcc ggagtacctg gcccctgagg tgctggagga caacgactac ggccgtgcag
     4021 tggactggtg ggggctgggc gtggtcatgt atgagatgat gtgtggccgc ctgcccttct
     4081 acaaccagga ccacgagaag ctgttcgagc tgatcctcat ggaggagatc cgcttcccgc
     4141 gcacactcgg ccctgaggcc aagtccctgc tctccgggct gctcaagaag gaccctacac
     4201 agaggctcgg tgggggctct gaggatgcca aggagatcat gcagcaccgg ttctttgcca
     4261 acatcgtgtg gcaggatgtg tatgagaaga agctgagccc acctttcaag ccccaggtca
     4321 cctctgagac tgacaccagg tatttcgatg aggagttcac agctcagatg atcaccatca
     4381 cgccgcctga tcaagatgac agcatggagt gtgtggacag tgagcggagg ccgcacttcc
     4441 cccagttctc ctactcagcc agtggcacag cctacccata cgacgtccca gactacgctt
     4501 gaaagggtgg gcgcgccgac ccagctttct tgtacaaagt ggttgatatc cagcacagtg
     4561 gcggccgctc gagtctagag ggcccgcggt tcgaaggtaa gcctatccct aaccctctcc
     4621 tcggtctcga ttctacgcgt accggttagt aatgagttta aacccgctga tcagcctcga
     4681 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     4741 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     4801 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     4861 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     4921 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaaataact
     4981 tcgtataatg tatgctatac gaagttatat cgtatgcatg ctttgcatac ttctgcctgc
     5041 tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagaa ttccttgaaa
     5101 tacatccaca ggtacacctc caattgactc aaatgatgtc aattagtcta tcagaagctt
     5161 ctaaagccat gacatcattt tctggaattt tccaagctgt ttaaaggcac agtcaactta
     5221 gtgtatgtaa acttctgacc cactggaatt gtgatacagt gaattataag tgaaataatc
     5281 tgtctgtaaa caattgttgg aaaaatgact tgtgtcatgc acaaagtaga tgtcctaact
     5341 gacttgccaa aactattgtt tgttaacaag aaatttgtgg agtagttgaa aaacgagttt
     5401 taatgactcc aacttaagtg tatgtaaact tccgacttca actgtagggt accgagctcg
     5461 aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     5521 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     5581 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt
     5641 ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc
     5701 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     5761 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     5821 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     5881 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     5941 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     6001 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     6061 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     6121 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     6181 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     6241 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     6301 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     6361 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     6421 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     6481 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     6541 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     6601 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     6661 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     6721 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     6781 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     6841 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     6901 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     6961 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     7021 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     7081 aaaggatctt c
//
